cover lass vant Ret Johannes ramp Start of Citation[ PUIM. Dekker|/PUJ[DP]1991|/DP]End of Citation Basic Bioreactor Design Riet, Klaas van 't. 0824784464 9780824784461 9780585157214 language: subject publication date: Ice: dde: subject: cover next page > next page >< previous page page_i Basic Bioreactor Design Klaas van 't Riet Johannes Tramper ‘Wageningen Agricultural University ‘Wageningen, The Netherlands i Mancet. Dex, Isc New York + Bastt Start of Citation[PUIM. Dekker[/PU][DP]1991|/DP]End of Citation < previous page page_i next page > Page i next page >< previous page page_ii Library of Congress Cataloging--in—Publication Data Riet, Klaas van't Basic bioreactor design / Klaas van 't Riet and Hans Tramper. p. cm Includes bibliographical references and index. ISBN 0--8247--8446--4 (acid-free paper) 1. Bioreactors----Design. I. Tramper, Hans II. Title. ‘TP248,25.B55R54 1991 660'62----de20 90--23465 cP ‘This book is printed on acid-free paper. Copyright © 1991 by MARCEL DEKKER, INC. All Rights Reserved next page > Page ii Neither this book nor any part may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, microfilming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher. MARCEL DEKKER, INC. 270 Madison Avenue, New York, New York 10016 Current printing (last digit): 109876 PRINTED IN THE UNITED STATES OF AMERICA Start of Citation[PUIM. Dekkerl/PU][DP]1991/DP]End of Citation < previous page page next page >< previous page page_i next page > Page i PREFACE Basic Bioreactor Design is based on the course material of the graduate course in biochemical engineering at Wageningen Agricultural University. This course has two objectives: 1 To provide the basic principles of reactor design 2 To select the relevant principles and data for practical process engineering purposes ‘These objectives determined the scope of material covered in this book. A limited number of reactor types as well as a limited number of phenomena are discussed. Yet we assume that the ones dealt with are enough to solve 95% of the problems and questions encountered in commercial fermentation. The book is intended for two groups of people: first, graduate students, to use as a textbook for learning the basic principles and methods of bioreactor design, and second, company engineers and biotechnologists, to use it as a handbook of fermenter design and engineering, Part of this book also originated from the time when Professor K. van 't Riet (then a Gist brocades employee) together with Professor J.J. Heijnen (then a Gist brocades employee, now with Delft ‘Technical University) cotaught a course on biochemical engineering with Dr. N.W.F. Kossen (then at Delft Technical University, now with Gist brocades). A substantial portion of Part Three originates from this lecture Series. ‘The authors wish to acknowledge Dr. N.W.F. Kossen and Prof. J.J. Hefnen for the contribution they made to this book, Prof. A. Prins is acknowledged for Chapter 12. Further, a number of the examples were contributed by Dr. L.E.S. Brink, Ir. C.D, de Goyer, Ir. M.H. Ziwvietering and Dr. P. Verlaan, Dr. R.G.J.M. van der Lans and Mr. B. Brandt made many useful suggestions and corrections. Also, we thank Mr. C. Rijpma and Mr. M. Schimmel for the artwork. Finally, layout and final preparation has been done by Mr. G. Heida and Ms. H.S. Wessels. Without their dedication this book never would have reached the final completion. KLAAS VAN 'TRIET HANS TRAMPER Start of Citation[ PUM. Dekker|/PU][DP]1991|/DP]End of Citation < previous page page_iii next page >< previous page page_v next page > Page v CONTENTS Preface ii Part One Introduction 1 3 Introduction 2 2 Balances 3 31 Yield 4 68 Kinetics 5 91 Stability 6 104 Flow, Flooding, Dispersion 7 122 Mec 8 136 Shear Part Three Reactor Engineering 9 183 Mixing 10 220 Hold-Up ul 236 Mass Transfer 12 274 Foam 13 293 Heat Transfer14 Power Consumption Part Four Reaction Engineering: Integration 15 Macrokineties: Particle Level 16 Apparent Stability 7 Macrokinetics: Reactor Level 18 Process Engineering Symbols Index Start of Citation[PUIM. Dekkerl/PU][DP]1991[/DP]End of Citation < previous page page_v 303 342 370 401 451 461 next page >PART | Introduction1 Introduction 1.1 Defining the subject Bioreactor design is an integral part of biotechnology, an area with rather loose and contested borders. A widely distributed, much advertised, but unfortunately wrong definition of biotechnology is: Microbiology Genetics Biochemi Engineering Chemistry Pharmacy Food Technology BIOTECHNOLOGY Biotechnology is not simply the sum of microbiology, try, engineering, etc.; no, it is the integration of these disciplines, and this involves quite a bit more than just simple addition. Integration and application are the keywords which can be found in most definitions of biotechnology (Fig. 1.1). Especially when designing bioreactors, integration of biological and engineering principles is essential. The bioreactor should be designed such that specific biological and technological demands of a process are met. Naturally, quality and price of the product are decisive for commercial realization. The aim of bioreactor design can thus be defined as “minimization of the costs of the pertinent product while retaining the desired quality, and this within the biological and technological constraints.” This does not mean & priori that minimizing the costs of the bioreactor also ‘means minimizing the costs of the integral process. This depends largely on the cost-determining part(s) of the process. If running the bioreactor is cost determining, then maximization of the overall volumetric productivity of 34 Chapter 1 ce siovocy LSB Psd (oNEERING SENTING DISCPLNES BIOTECHNOLOGY 1 t | i eDical, jnenostics ‘APPLICATION AREAS Fig. 1.1. Biotechnology: application oriented integration of biodiscipines and engineering. the bioreactor is, in general, the rational approach. If, on the other hand, the downstream processing is cost determining, then maximization of the product concentration in the bioreactor is, in general, the rational thing to do, However, here again integration is the keyword. Bioreactor design should be an integral part of the overall process design.Introduction 5 The words bioreactor, biocatalyst and product have been used in general terms. In the following sections of this chapter the bioreactor will be defined, though still in general terms, with respect to reactor concepts and types and to tools in bioreactor design. The meaning of product is obvious th the annotation that biomass can be the desired product too. In this book biocatalyst means either an enzyme, an enzyme complex, a cell organelle or 1a whole cell. The latter can be growing or nongrowing, viable or nonviable, etc. Furthermore, a biocatalyst can be free or immobilized, which has far-reaching consequences not only with respect to mass transfer, but sometimes also for the physiology of viable cells. Integration of mass transfer and biokinetics is essential in the description (microkinetics) of ‘mobilized biocatalysts. The source of biocatalysts can be of either microbial, plant or animal origin and examples of all three are used in this book. 1.2 Productivity and product concentration 1.2.1 Overall volumetric productivity Overall volumetric productivity Q, (mol m-S 5-4) (itis also common to use a yearly basis) is the average production capacity per unit volume and time of the bioreactor. The overall volumetric productivity is confined on the one hand by physical constraints such as mass and heat transfer, and on the other hand by biocatalyst concentration C, (mol m-8) and activity of the bio- catalyst, expressed as substrate consumption rate -ri (mol mS s°}). Maximization of the overall volumetric productivity of the bioreactor in principle means minimization of the costs of investment, because one can suffice with smaller equipment. It usually also means lower operating costs. In general, it means too that it is desired to operate the bioreactor as close as possible to the physical constraints, the horizontal dotted line in Fig. 1.2. ‘This physical limitation is the result of mass and heat transfer limitations, which are stoichiometrically related to product formation. The vertical dotted line in Fig. 1.2 symbolizes the limitation which is a consequence of the fact that the concentration of the biocatalyst is bound to certain defined limits, for instance solubility in case of isolated enzymes and “space” in case of suspended cells. Fig. 1.2 also shows that the biocatalyst should have a minimum specific activity to be able to operate the bioreactor close to its physical ceiling. 1.2.2 Overall blocatalyst productivity In addition to limitations by mass and heat transfer and concentration of biocatalyst, the overall volumetric productivity of the bioreactor is deter-6 Chapter 1 volumetric productivity Qp Fig. 1.2 Constraints of overall volumetric Productivity. (Adapted from biocatalyst cone. Cx Gooney, 1983) mined by the overall productivity of the biocatalyst, Pr yx (-), defined as the total moles of product which are produced by 1 mol of biocatalyst uring its operational lifetime t, (s). Pr p, is related to the specific product production rate q, (3-4) (moles of product produced per mol of biocatalyst per second) as Pre * f aya 7 -frnBa oO ou ‘ ‘ * The definition of ¥'$, (-), the overall yield of product on substrate (total moles of product produced per total mol of substrate), leads to 7 apCxdt (mol m$s2) 12a f -Yertat (mol ms) 1.2b °INTRODUCTION 7 ‘The time needed to empty, clean, refill, restart, etc., the bioreactor between two operations is the so-called down-time, which is symbolized by {4 (s). In case ty is relevant it can be introduced in Eq. (1.2) by replacing 1/t,, preceding the integral, by 1/(t,+ 14). In addition to the molar productivity used above, the mass productivity (kg product instead of mol) is also quite commonly used in engineering (conversion from one to the other by means of the molecular weights). It is also common practice to use hour, day or year as unit of time, ‘The search for and the development of a useful biocatalyst with a suitable yield, specific activity and stability is in the beginning the task of micro- biologists, biochemists, molecular biologists, protein engineers, etc. How- ever, especially with respect to stability, the process engineer also has means available, among others immobilization, to improve the stabilit biocatalysts. This is covered in this book. 1.2.3 Product concentration The effect of the composition of the product stream leaving the bioreactor ‘on the costs of the downstream processing is large. Therefore, it is essential to take this into account when designing the bioreactor. This often means in practice that the bioreactor is designed such that the concentration of product is as high as possible, The end concentration of product C (mol ms) in the bioreactor depends on ‘and the residence time the bioreactor. For a batch reactor, with ‘fy (s) as the time that the batch lasts, this leads to chs vn f ria (mot m’) 13 and for a continuous reactor with a liquid throughflow F, (m* s-) and a volume V (m?); Se | (mol m-$) 1.4 Concentration of product is especially a key-parameter when the down- stream processing is the cost-determining part of the integral process. Product recovery is often a laborious and expensive operation, especially when diluted aqueous solutions are involved such as we usually encounter in biotechnology. However, it has become clear that the aqueous reaction medium, which was for a long time supposed to be essential for biocatalysis, Gan be replaced to large extent by «suitable organic solvent (Laane eta 1987).1.3 Bloreactor types 1.3.1 The stirred vessel In Fig. 1.3 a schematic view of a stirred vessel is given. The vessel is cylindrical with a height H, (m) and a diameter T, (m). Usually H, is equal to or greater than 27. It is equipped with a stirrer in the lower compartment. This stirrer is mounted near the bottom usually at a distance equal to the stirrer diameter. At a lower position the stirrer and bottom interact leading to a decrease in power consumption. At a higher position liquid circulation problems can occur because at increased gas flow rate in case of aeration the bubbles will not be recirculated in the lower compart- ‘ment. Sometimes the upper compartment(s) are also equipped with a stirrer. The vessel is equipped with baffles to prevent rotation of the contents as a whole. For aeration an air sparger is mounted below the stirrer. For mass transfer its construction is generally not relevant, so it is chosen on the basis of sterility and cleaning criteria. Fig. 1.4 shows a number of stirrers that are used. It will be shown in Chapter 11 that the stirrer is needed to provide a certain level of power input needed Fig. 13 Schematic representation of & stirred formenterIntroduction. 9 Bidt six-blade six-blade 45° paddle MIG-agitator disc turbine pitched turbine agitator Fig. 1.4 Schematic representation of a numberof sirers. (Adapted from Zlokarrik, 1972.) for aeration purposes. Therefore, the turbine stirrer, being easy to construct and having a high power number, is the most widely used. The other types are less intensively applied. A detailed description of all types of stirrers can be found in Zlokarnik (1972). Special design considerations like stirrer drives and sealings are not dealt with here, Also self-aerating stirrers are not discussed. Information about them can be found in Sittig (1983). 1.3.2 The bubble colu ‘A schematic representation of this simple reactor is given in Fig. 1.5. Usually it is H, 2 27,. At the bottom a sparger is mounted. To prevent too heterogeneous flow patterns in the lower compartment, the sparger nozzles have to be distributed over the cross section of the bottom. There- fore, one ring or a small number of parallel pipes or a starlike construction of pipes is commonly used. In the pipes holes are drilled. In the chapter on ‘mass transfer it is shown that complicated spargers or very small holes merely have disadvantages for most appl 1.3.3 The alr lift ‘The air lift consists of two pipes, interconnected at top and bottom. In one of the pipes (the riser) air is sparged at the bottom. The air rises and escapes at the top. Therefore, under most circumstances there is no air present in the other pipe (the downcomer). The density difference between riser and10 Chapter 1 Fig. 1.8 Schematic representation of a ‘bubble column reactor. downcomer causes an intensive liquid circulation. Two designs can be used, i.e., the internal (Fig, 1.64) and the external loop reactor (Fig. 1.6B). When an internal loop reactor is built underground, we refer to this as a deep shaft. Volumes can be up to thousands of m’.'H, generally is much larger than T, , usually of the order of 10 T,, but for the deep shaft up to 100 7, . downcomer —+ riser + air Fig. 16 Schematicrepresentation ofthe air. A. Intemalloop reactor. B. Externalloop reactor.INTRODUCTION 1" 1.3.4 The packed bed ‘The packed bed is very simple in nature and differs largely from the other three types. It consists of a tubular pipe, packed with the biocatalyst par- ticles. It can be operated in the upflow or downflow mode, i.e, the reaction medium is introduced either at the bottom or the top of the reactor. 1.3.5 Discussion of reactor types ‘The four basic reactor types discussed in this book are the ones described above. The difference between the packed bed and the other three is so great that the choice usually is straightforward. On the basis of detailed mixing, mass transfer and cost calculations a rational choice can be made between stirred vessel, bubble column and air lift. Many times, however, not needed and the following general rules suffice. - Mixing and mass transfer do not differ very much between the three types. In the air lift the flow is controlled, which enables the intro- duction of a controlled substrate addition at more than one place. - For mass transfer the maximum attainable value is higher for stirred vessels, because of the larger power that can be introduced with the stirrer. For air lift and bubble column mass transfer collapses above a viscosity of about 0.1 Ns m2, = At increasing scale mechanical problems can occur in the stirred vessel because of the large power values of the stirrer motor. Based on these three reasons the follo 18 rules of thumb apply: = In those cases where viscosities can rise above 0.1 Ns m-? (mycelial, biopolymer fermentations) a stirred vessel is chosen because air lift and bubble column will = In those cases where flexibility in viscosity and mass transfer is needed (pilot plant) a stirred vessel is chosen because air lift and bubble column cannot offer this flexibility, = In low viscosity fermentations at large scale (50-500 m8) a bubble col- umn is chosen because it is the cheapest fermenter. = In low viscosity fermentations at very large scale (200-10,000 m3) an ai lift is chosen because it permits local and controlled substrate addition, (The stirred fermenter would offer immense mechanical problems at V > 500 m? because the stirrer power P, could easily rise above 1 MW. - Viscous fermentations cannot be scaled up to scales > ~ 500 m* because the stirred fermenters that are needed for these cases will offer mech- anical problems ( P, > 1 MW).